Method for obtaining differentiated cells and/or differentiated cell products from undifferentiated cell and a method of perfusion culture

Abstract

Disclosed is a method for obtaining differentiated cells and/or differentiated cell products, including the steps of: introducing undifferentiated cells into a perfused organ or living tissue; subjecting the undifferentiated cells introduced to perfusion culture together with the organ or living tissue so as to allow the undifferentiated cells to differentiate, thereby obtaining differentiated cells and/or differentiated cell products; collecting a perfusion culture solution that contains the resulting differentiated cells and/or differentiated cell products; and obtaining the differentiated cells and/or differentiated cell products contained in the collected perfusion culture solution.

What is claimed is: 
     
       1. A method for obtaining differentiated cells and/or differentiated cell products, comprising the steps of:
 introducing undifferentiated cells into a perfused living organ or a perfused living tissue, wherein the living organ or living tissue is a living organ or living tissue extracted from an animal other than humans; 
 subjecting the undifferentiated cells to perfusion culture together with the living organ or living tissue so as to allow the undifferentiated cells to differentiate; 
 collecting a perfusion culture solution comprising the resulting differentiated cells and/or differentiated cell products, wherein the resulting differentiated cells are cells which were differentiated from the undifferentiated cells in the living organ or living tissue in the subjecting step, and wherein the differentiated cell products are products which were produced by the cells differentiated from the undifferentiated cells in the living organ or living tissue in the subjecting step; and 
 obtaining the differentiated cells and/or differentiated cell products comprised in the collected perfusion culture solution, 
 wherein the differentiated cells are allogeneic to the living organ or living tissue. 
 
     
     
       2. The method according to  claim 1 , wherein the undifferentiated cells are human-derived cells. 
     
     
       3. The method according to  claim 1 , wherein the undifferentiated cells are stem cells or precursor cells. 
     
     
       4. The method according to  claim 3 , wherein the stem cells are at least one selected from the group consisting of ES cells, cloned ES cells, mesenchymal stem cells, and iPS cells. 
     
     
       5. The method according to  claim 3 , wherein the precursor cells are at least one selected from the group consisting of platelet precursor cells, liver precursor cells, heart precursor cells, and neuronal precursor cells. 
     
     
       6. The method according to  claim 5 , wherein the platelet precursor cells are at least one selected from the group consisting of megakaryocyte precursor cells, megakaryoblasts, and promegakaryocytes. 
     
     
       7. The method according to  claim 5 , wherein the liver precursor cells are at least one selected from the group consisting of hepatoblasts, hepatic precursor cells, hepatic stellate precursor cells, hepatic stem/precursor cells, vascular endothelial precursor cells from liver, and mesothelial precursor cells from liver. 
     
     
       8. The method according to  claim 5 , wherein the heart precursor cells are cardiac muscle precursor cells or vascular endothelial precursor cells from heart. 
     
     
       9. The method according to  claim 5 , wherein the neuronal precursor cells are at least one selected from the group consisting of neuron precursor cells, glial precursor cells, and vascular endothelial precursor cells from central nervous system. 
     
     
       10. The method according to  claim 1 , wherein the living organ or living tissue is at least one selected from the group consisting of spleen, liver, pancreas, heart, lung, kidney, and bone. 
     
     
       11. The method according to  claim 10 , wherein the living organ or living tissue is spleen, and the differentiated cells are mature megakaryocytes and/or osteoblasts. 
     
     
       12. The method according to  claim 10 , wherein the living organ or living tissue is bone, the differentiated cells are mature megakaryocytes, and the differentiated cell products are platelets. 
     
     
       13. The method according to  claim 1 , wherein the introduction step is performed by adding the undifferentiated cells to a perfusate. 
     
     
       14. The method according to  claim 1 , further comprising the step of determining whether or not the introduction step is performed by evaluating a storage state of the living organ before the introduction step. 
     
     
       15. The method according to  claim 14 , wherein in the determination step the storage state of the living organ is evaluated by measuring at least one selected from lactate dehydrogenase in the perfusate, lactic acid in the perfusate, perfusion pressure, and dissolved oxygen. 
     
     
       16. A method of perfusion culture with a perfusion culture system using a perfused living organ or a perfused living tissue, comprising the steps of:
 measuring an activity of lactate dehydrogenase of a perfusate on a day the living organ or the living tissue is extracted, the extracted living organ or living tissue being placed in the perfusate; 
 introducing undifferentiated cells into the perfused living organ or the perfused living tissue; 
 subjecting the undifferentiated cells to perfusion culture together with the living organ or living tissue so as to allow the undifferentiated cells to differentiate; 
 collecting a perfusion culture solution comprising the resulting differentiated cells and/or differentiated cell products, wherein the resulting differentiated cells are cells which were differentiated from the undifferentiated cells in the living organ or living tissue in the subjecting step, and wherein the differentiated cell products are products which were produced by the cells differentiated from the undifferentiated cells in the living organ or living tissue in the subjecting step; and 
 obtaining the differentiated cells and/or differentiated cell products comprised in the collected perfusion culture solution, 
 wherein the differentiated cells are allogeneic to the living organ or living tissue. 
 
     
     
       17. A method of perfusion culture with a perfusion culture system using a perfused living organ or a perfused living tissue, comprising the steps of:
 introducing platelet precursor cells into a perfused living organ or a perfused living tissue, the living organ or the living tissue being extracted from an animal other than humans and placed in a perfusion culture solution in a container of the perfusion culture system, the living organ being selected from the group consisting spleen, liver, pancreas, lung and kidney, the living tissue being a bone; 
 subjecting the platelet precursor cells to perfusion culture together with the living organ or living tissue in the perfusion culture solution in the container so as to allow the undifferentiated cells to differentiate into megakaryocytes producing platelets; 
 collecting the perfusion culture solution comprising the platelets from the living organ or living tissue, wherein the platelets are products which were produced by the megakaryocytes differentiated from the platelet precursor cells in the subjecting step; and 
 obtaining the platelets comprised in the collected perfusion culture solution.